Effect Of Myeloid MRD-L Patients CD34 ~ + Cells Derived DC During The Induction Of IL-12 Modified Biejia Soup

Objective:The fundamental reason of recurrence after of Acute leukemia is minimal residual leukemia relapse, that body fails to effectively remove the residual leukemia cells, because tumor immunity can not play as normal.the key link of this process is the leukemia cells as tumor antigen recognition, presenting.the most powerful presenting cells of the priming antigen primary immune is dendritic cells。the number of dendritic cells and the ability of antigen presenting play the key role in starting tumor immune period effectively of acute leukemia. how to Increase the number of mature dendritic cells (DC),and how environment affect the number of dendritic cells and mature state has become the focus of many scholars to study. In this experiment, the method of cell culture in vitro, containing different doses of Jiawei Qinghao Biejia Decoction of the animal serum and cytokines,We culture from the complete remission of acute myeloid leukemia (AML-CR) during the extraction and separation of CD34+cells in patients with bone marrow, which induced into dendritic cells (DC), to observe the effects of different concentrations of serum at different stages of dendritic cell maturation, and IL-12in different stages of the content change of serum, and study the treatment of minimal residual leukemia from the traditional Chinese medical science point of indirect immunofluorescence of Jiawei Qinghao Biejia decoction, could induce DC maturation and secretion of IL-12effects, and IL-12and DC cytokines in the between.Methods:Draw30ml of blood samples from AML-CR bone marrow, obtained bone marrow mononuclear cells by Ficoll density gradient centrifugation (BMNC),and then by immunomagnetic positive selection method for extraction of high purity CD34+cells. The CD34+cytokines (SCF, TPO, FLT-3) for in vitro amplification, amplification of6-9days after reaching a certain number of cell culture plate with6holes were divided into5groups: culture.5groups with10%FCS RPMI1640as the base fluid, with traditional Chinese medicine and cytokines in different concentrations (GM-CSF, IL-4, IFN-α, TNF-α) are divided into high, low, traditional Chinese medicine groups (concentration in serum and animal cell factor containing different concentrations of traditional Chinese Medicine), the blank serum group (feed with normal serum and animal cell factor a), cell factor group (no animal serum, only the cell factor). Training in the same environment for about9days, change liquid every2～3days, and give close observation of growth and morphology of cells in culture, zeroth,6,9were detected by ELISA IL-12in the supernatant serum in the expression of mature cells at the same time,, flow cytometry was used to detect the cell surface molecules CD80, CD83, CD86, HLA-DR, CDla Results:1. The morphology of DC:Observation under inverted microscope, zeroth days just separated CD34+cells are large and round, suspended in the cell culture medium;In the third day the cells form a small set of scattered in the suspended in liquid culture, volume become larger and shape become Irregular. In the sixth day, Cell colony was significantly increased, the cell morphology was irregular, a part of the cells were short spikes;In the seventh day, cell volume increased further, presentd a short branch, the number of cells increased;In the ninth day, DC formed with typical morphology, The cell periphery tree mutation elongated,and the number of cells increased. Groups with serum and cytokines which containing traditional Chinese medicine, their DC morphology are more typical than cytokine group and blank serum group. There is no significant difference in morphology between the traditional Chinese medicine groups; the number of DC in the traditional Chinese medicine groups is slightly higher than the other groups.2. Detection of IL-12content changes:The contents of the traditional Chinese medicine combined with cytokines in the ninth day, and sixth day were higher than those in zeroth days, Chinese medicine group was higher than that of cell factor group and blank serum group, the content of medium dose of traditional Chinese medicine group was higher than that of other groups. JWZ group inthe ninth day was significantly higher than that in the sixth day (P<0.01).3. All groups have the expression of dendritic cell immune phenotype. Groups containing different concentration of traditional Chinese medicine compared with the blank group the expression of CD80, CD83, CD86, and they showed no statistical significance. In the middle and low concentration of traditional Chinese medicine serum group, CDla expression was statistically significant compared with the other group. In the groups of different concentration of traditional Chinese medicine serum, the expression of HLA-DR cytokine group had statistical significance compared with the blank serum group.Conclusion:Jiawei Qinghao Biejia Tang Xueqing with combined cytokines GM-CSF, IL-4, IFN-α,TNF-α,induced culture dendritic cells and promoted the growth, differentiation and maturation of DC better than only using cytokine In vitro. it may relate to the Jiawei Qinghao Biejia decoction which affect the cell growth mature environment, such as affecting the content of some factors, so as to promote the maturation of DC, and can promote the secretion of IL-12dendritic cells, increase the effects of antitumor.